Physical activity is a critical component of obesity prevention, but few interventions targeting early childhood have been described. The Active Early guide was designed to increase physical activity in early care and education (ECE) settings. The purpose of Active Early 2.0 was to evaluate the effectiveness of Active Early along with provider training, microgrant support, and technical assistance over 2 years (2012-2014) to increase physical activity and related behaviors (e.g., nutrition) in settings serving a high proportion of children from underserved groups in recognition of significant disparities in obesity and challenges meeting physical activity recommendations in low-resource settings. The physical activity and nutrition environment were assessed before and after the intervention in 15 ECE settings in Wisconsin using the Environment and Policy Observation Assessment tool, and interviews were conducted with providers and technical consultants. There was no significant change in Total Physical Activity Score or any EPAO subscale over the intervention period; however, significant improvements in the Total Nutrition Score and the several Nutrition subscales were observed. Additionally, the percentage of sites with written activity policies significantly increased. Overall minutes of teacher-led physical activity increased to 61.5 ± 29.0 min (p < 0.05). Interviews identified key benefits to children (i.e., more energy, better rest, improved behavior) and significant barriers, most notably care provider and child turnover and low parent engagement. Moderate policy and environmental improvements in physical activity and nutrition were achieved with this intervention, but more work is needed to understand and address barriers and to support sustained changes in lower-resource ECE settings.